miRNA-148a 在膀胱癌组织中的表达及生物信息学分析

Expression of miRNA-148a in Bladder Carcinoma Tissues and Its Bioinformatics Analysis

目的：探索 miRNA-148a 在膀胱癌发生发展中的作用。方法收集35例膀胱癌组织和16例非癌组织，采用荧光定量 PCR 方法检测 miRNA-148a 的相对表达量，并分析其与临床特征的相关性；采用生物信息学分析，预测 miRNA-148a的靶基因和转录因子，构建 TF-miRNA-148a-靶基因调控网络图，并对其靶基因进行 GO 富集和 KEGG Pathway 分析。结果miRNA-148a 在膀胱癌中的相对表达量（0.0008±0.0002）明显低于非癌组织（0.0021±0.0005）（t＝2.46，P ＜0.05），其相对表达量在不同性别、年龄、病理分级、临床分期、淋巴结转移组的差异无统计学意义（P ＞0.05）。3个软件都预测到的 miRNA-148a 靶基因268个；预测到 miRNA-148a 的转录因子60个，结合评分＞80的结合位点657个；对268个靶基因进行 GO 富集分析，发现其靶基因主要涉及神经细胞分化、发育、增殖、mRNA 合成，蛋白连接等众多的生物学过程（P ＜0.001，相对于背景具有统计意义）；KEGG Pathway 分析发现268个靶基因参与 p53，恶性肿瘤、前列腺癌、PI3K-AKT 等信号通路（P ＜0.05，相对于背景具有统计意义）；根据构建的 TF-miRNA-148a-靶基因调控网络图，挖掘出可能调控 miRNA-148a 的转录因子有 SP1，ESR1，AP1，MYC，BRCA1等；可能受 miRNA-148a 调控的基因有 IGF1，P27 kip1，NCOA1，PTEN， SERPINE1等。结论miRNA-148a 在膀胱癌中表达下调，可能参与膀胱癌的发生发展，生物信息学分析为后续研究提供一定的思路。

Objective To explore the role of miRNA-148a in bladder tumorous development and progression.Methods Ex-pression of miRNA-148a was assessed in 35 bladder carcinoma tissues and 16 non-carcinoma tissues by fluorescence quanti-tative real time PCR,and correlation with clinical features was evaluated.Target genes and transcription factors of miRNA-148a were predicted using bioinformatic analysis,then TF-miRNA-148a-target genes network diagram was built and the tar-get genes was analyzed of gene ontology enrichment and KEGG pathway.Results Expression of miRNA-148a was lower in bladder carcinoma tissues than in non-carcinoma tissues（0.000 8±0.000 2 vs 0.002 1±0.000 5）（t=2.46,P 〈0.05）,but its expression was no statistical significance in different groups of gender,age,pathological classification,clinical stage, lymph node metastasis （P 〉0.05）.268 target genes of miRNA-148a were predicted by three softwares at the same time,60 transcription factors were predicted and the binding sites with combination scroes above 80 was 657.The target genes of miRNA-148a was enriched in many biological processes,such as neuron differentiation,generation of neurons,neuron projec-tion development,cytoplasmic mRNA processing body,cytoplasm（P 〈0.001）.They also participated in p53 signaling path-way,proteoglycans in cancer-homo sapiens,pathways in cancer,prostate cancer,protein processing in endoplasmic reticulum, focal adhesion and so on（P 〈0.05）.According to TF-miRNA-148a-target genes network diagram,miRNA-148a was regula-ted by SP1,ESR1,AP1,MYC and BRCA1,genes of IGF1,P27kip1 ,NCOA1,PTEN,SERPINE1 might be regulated by miR-NA-148a.Conclusion miRNA-148a which was significantly down-regulation in bladder carcinoma tissues may be participate in bladder tumorous development and progression,bioinformatics analysis provides some ideas for further research.